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Background: The purpose of this study was to find out the incidence and characteristics of H. influenzae type b invasive disease (HibID) in Catalonia, Spain., Material and Methods: An active surveillance of H. influenzae isolated from normally sterile sites was carried out during 1996. Microbiology laboratories of hospitals of Catalonia were periodically contacted by telephone. The serotype of all the strains was studied., Results: The incidence of H. influenzae invasive disease (HIID) was 7.1 per 100,000 in children under 5 years and 1.0 per 100,000 in those over 5 years. The incidence of serotype b was 6.4 per 100,000 children under 5 years and 0.2 above this age. Only three strains belonged to types other than b (d, e and f)., Conclusions: The incidence of HIbID is uncommon in Catalonia, lower than that registered in the prevaccine era in other countries and regions of the same geographical area.

Background: Erythema induratum of Bazin (BEI), is included in the group of cutaneous granulomatous lobulillar panniculitis. The aethiopathogenic association between EI and tuberculosis can not rely on the clinicohistological features of these panniculitis and M. tuberculosis has never been isolated from BEI lesions. Detection of the mycobacterial DNA by PCR on cutaneous biopsy samples would allow to confirm this association., Patients and Methods: Fourteen patients with clinical BEI were chosen retrospectively. Seventeen lesional biopsy samples were obtained, stained with the Kinyoun carbolfuchsin acid-fast technique and haematoxylin and eosin and tested by PCR. A fragment of the IS6110 insertion sequence specific of M. tuberculosis was amplified and confirmed by digestion with Sal I restriction endonuclease. The efficiency of the procedure, the presence of inhibitory substances and the preservation of DNA were checked by PCR of the beta-actin gene., Results: M. tuberculosis DNA was detected in 12 of the 17 samples tested (70.5%) which corresponded to 10 of the 14 patients (71.4%). According to beta-actin PCR results, the rate of extracted DNA was inadequate on four of the five negative biopsies., Conclusions: The results of these series suggest the probable involvement of M. tuberculosis on the BEI pathogenesis and give support to the usefulness of the PCR in the diagnosis of this pathology concerning the need of specific treatment.

Hemorrhagic colitis is an enteritis caused by verotoxigenic strains of Escherichia coli. Conventional diagnosis requires the identification of the microorganism and the demonstration of verotoxin production. The determination of toxigenicity in isolated strains and in direct stool samples by the polymerase chain reaction (PCR) technique may simplify the diagnosis. Conventional coprocultures were performed for the detection of verotoxigenic E. coli O157:H7 from three stool samples of a patient with hemorrhagic colitis and hemolytic-uremic syndrome. The production of verotoxin was determined by cell culture and the presence of VT1 and VT2 genomic sequences by PCR. Likewise, the latter technique was applied to a direct stool sample for detection of the verotoxin codiying genes. The specificity of the amplified sequences was confirmed by enzyme restriction digestion. Escherichia coli O157:H7 was isolated in two of the three samples studied. The strains were toxigenic in the cell culture test at titers higher than 1/500 and PCR showed an amplified band of 479 pb corresponding to the VT2 codifying gene. The digestion of amplified sequences with the EcoRV enzyme led to two bands of 390 and 89 pb confirming the specificity of the results. One of the two stool samples studied directly by PCR was positive for VT2 with the result being obtained 48 hours after arrival to the laboratory. The preliminary results of this study give support to the usefulness of the polymerase chain reaction technique in the detection of verotoxin from isolated strains of Escherichia coli and in direct stool samples.